EP3486324B1 - Method for preparing amines from carbonyl compounds by transaminase reaction under salt precipitation - Google Patents

Method for preparing amines from carbonyl compounds by transaminase reaction under salt precipitation Download PDF

Info

Publication number
EP3486324B1
EP3486324B1 EP17202282.4A EP17202282A EP3486324B1 EP 3486324 B1 EP3486324 B1 EP 3486324B1 EP 17202282 A EP17202282 A EP 17202282A EP 3486324 B1 EP3486324 B1 EP 3486324B1
Authority
EP
European Patent Office
Prior art keywords
alkyl
unbranched
branched
group
hydrogen atom
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
EP17202282.4A
Other languages
German (de)
English (en)
French (fr)
Other versions
EP3486324A1 (en
Inventor
Jan von Langermann
Dennis Hülsewede
Marco Tänzler
Ulf Menyes
Philipp Süß
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Enzymicals AG
Universitaet Rostock
Original Assignee
Enzymicals AG
Universitaet Rostock
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority to DK17202282.4T priority Critical patent/DK3486324T3/da
Application filed by Enzymicals AG, Universitaet Rostock filed Critical Enzymicals AG
Priority to EP17202282.4A priority patent/EP3486324B1/en
Priority to ES17202282T priority patent/ES2816102T3/es
Priority to PCT/EP2018/081517 priority patent/WO2019096973A1/en
Priority to KR1020207017054A priority patent/KR102554463B1/ko
Priority to JP2020545446A priority patent/JP7250807B2/ja
Priority to US16/764,476 priority patent/US12018303B2/en
Priority to CN201880074226.2A priority patent/CN111465699B/zh
Publication of EP3486324A1 publication Critical patent/EP3486324A1/en
Application granted granted Critical
Publication of EP3486324B1 publication Critical patent/EP3486324B1/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C12P13/001—Amines; Imines
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P13/00—Preparation of nitrogen-containing organic compounds
    • C12P13/005—Amino acids other than alpha- or beta amino acids, e.g. gamma amino acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C211/00—Compounds containing amino groups bound to a carbon skeleton
    • C07C211/62—Quaternary ammonium compounds
    • C07C211/63—Quaternary ammonium compounds having quaternised nitrogen atoms bound to acyclic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C57/00—Unsaturated compounds having carboxyl groups bound to acyclic carbon atoms
    • C07C57/30—Unsaturated compounds having carboxyl groups bound to acyclic carbon atoms containing six-membered aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1096—Transferases (2.) transferring nitrogenous groups (2.6)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids

Definitions

  • the present invention relates to a method for preparing an amino salt compound comprising: i) providing a carbonyl compound; ii) reacting the carbonyl compound provided according to (i) in the presence of a transaminase with ii-a) at least one primary amine; and ii-b) at least one carboxylic acid; thereby obtaining a mixture comprising an at least partially crystallized amino salt compound comprising an amino cation and a carboxylate anion based on the at least one carboxylic acid added according to (ii-b).
  • the transamination-reaction can be carried out as a kinetic resolution of a racemic amine or an asymmetric synthesis from the respective prochiral ketone. Due to a maximum yield of 100% the asymmetric synthesis is in theory preferred, especially if a catalyst with high enantioselectivity is used.
  • thermodynamic limitations and certain product inhibitions tend to limit the applicability of transaminases in asymmetric synthesis, which needs to be overcome for synthetic purposes.
  • complex (co-)product removal techniques are currently considered, e.g. enzymatic cascades, membrane processes and non-catalytic side reactions. Such techniques unfortunately always increase (in general) overall complexity of the biocatalytic reaction systems, require additional or tailor-made co-substrates and generate further by-products.
  • the object of the present invention was therefore the provision of a method for the preparation of a desired product amine via biotransformation, especially using a transaminase, which overcomes the above-mentioned drawbacks.
  • Enzymes referred to in the following are known from e.g. WO 2011/026556 (present SEQ ID NOS 1-4 and enzymes closely related to SEQ ID NOS 5-6) and UniParc - UPI0009F6A547 (present SEQ 6).
  • amino salt compound comprises cation and anion as described above.
  • the amino salt compound can be present as pure salt compound (unsolvated, anhydrated) or as solvate or hydrate or mixture thereof, wherein hydrate includes hemihydrate, monohydrate and polyhydrate and solvate includes hemisolvate, monosolvate and polysolvate.
  • solvate includes hemisolvate, monosolvate and polysolvate.
  • At least partially crystallized means that at least 50%, preferably at least 60%, more preferably at least 70%, more preferably at least 80%, more preferably at least 90% of the amine salt compound precipitate in crystalline form.
  • hetero means that heteroatom(s) are present, which are selected from the group consisting of nitrogen atom, sulphur atom and oxygen atom, if not otherwise indicated.
  • the heteroatom(s) is/are present as members of the respective chain or ring structure.
  • R 1a, 2a are as defined above. It has to be noted that neither R 1a nor R 2a is a substituent which comprises a charge, i.e., substituents having a charge are excluded as R 1a and as R 2a .
  • R 1a and also R 2a are no carboxyl/carboxylate, no phosphate, no sulfonate.
  • the combination of R 1 , R 1a being a perfluorinated alkyl is excluded.
  • R 2 and R 2a i.e. the combination of R 2 and R 2a being a perfluorinated alkyl is excluded as well.
  • neither the combination of R 1 and R 1a nor the combination of R 2 and R 2a is a perfluorinated alkyl.
  • R 1 and R 2 together form a C3-C10-cycloalkyl or a C3-C10-cycloalkenyl
  • the number of C atoms indicated includes the carbonyl C atom situated between R 1 and R 2 .
  • Scheme 1 shows the method based on the exemplarily product amine 1-phenylethylamine.
  • the product amine 2 is herein selectively crystallized from solution as a barely soluble amine salt 6, while all other reactants, especially the applied donor amine (isopropyl amine, 3), remains in solution.
  • This in situ-product crystallization (ISPC) continuously removes the desired product amine from solution and thus yields an equilibrium displacement towards the products.
  • the counter ion here shown as a carboxylate
  • a stoichiometric use of the carboxylate in comparison to the applied amines is not required (see below).
  • a specific counter ion i.e. a specific carboxylic acid
  • the donor amine salt is not crystallized.
  • most commonly used amine salts, especially amine halide salts show very high solubilities in aqueous solutions and thus are not applicable for such an ISPC concept.
  • a preferred embodiment of the method is that the amino salt compound obtained according to (ii) has a solubility in water at pH 7 which is smaller than the solubility in water of the at least one primary amine added according to (ii-a).
  • a further preferred embodiment of the method is that the amino salt compound obtained according to (ii) has a solubility in water at pH 7 ⁇ 30 mmol/l, preferably ⁇ 25 mmol/l, more preferably ⁇ 10 mmol/l.
  • the solubility difference (delta Sol. ) between the solubility of the salt of the primary amine and the solubility of the amino salt compound is at least 10 mmol/l.
  • the solubility of the salt of the primary amine (sol. primary amine salt ) is higher than the solubility of the amino salt compound (sol. amino salt compound ), i.e.: sol. primary amine salt > sol. amino salt compound .
  • the used transaminase has to tolerate the required concentration of the chosen compound.
  • the formed salt has to be stable under process conditions and should be easily recovered from the reaction mixture.
  • step (ii) the carbonyl compound provided according to (i) is reacted in the presence of a transaminase with ii-a) at least one primary amine; and ii-b) at least one carboxylic acid.
  • the at least one carboxylic acid according to (ii-b) is used in its protonated form or in deprotonated form with a suitable counter cation.
  • the above-described straight-forward screening approach resulted in the finding, that the at least one carboxylic acid according to (ii-b) is a carboxylic acid of general formula (III) wherein n is zero or 1; the residues R 3 and R 4 are both phenyl or together form a phenyl ring, wherein each phenyl ring has at least one further substituent selected from the group consisting of hydrogen atom, halogen atom, preferably chloro, and nitro group; and the residue R 5 is hydrogen atom or methyl or absent if R 3 and R 4 together form a phenyl ring.
  • the straight-forward screening approach especially resulted in two benzylbenzene-based acids and three benzoic acid derivatives that matched the above mentioned criteria (as the respective carboxylate ions at pH 7.5)
  • a preferred embodiment of the method relates to the at least one carboxylic acid according to (ii-b) being a carboxylic acid selected from the group consisting of diphenylacetic acid (DPAA), 2,2-diphenylpropionic acid (2DPPA), 3,3-diphenylpropionic acid (3DPPA), 3,4-dichloro-benzoic acid (34CA), 3,4-dinitro-benzoic acid (34NA) and 4-chloro-3-nitro-benzoic acid (43CNA).
  • DPAA diphenylacetic acid
  • 2DPPA 2,2-diphenylpropionic acid
  • 3DPPA 3,3-diphenylpropionic acid
  • 34CA 3,4-dichloro-benzoic acid
  • 34NA 3,4-dinitro-benzoic acid
  • 4-chloro-3-nitro-benzoic acid 43CNA
  • the at least one carboxylic acid according to (ii-b) is selected from the group consisting of diphenylacetic acid (DPAA), 2,2-diphenylpropionic acid (2DPPA) and 3,3-diphenylpropionic acid (3DPPA), more preferably 2,2-diphenylpropionic acid (2DPPA) or 3,3-diphenylpropionic acid (3DPPA), more preferably at least 3DPPA.
  • DPAA diphenylacetic acid
  • 2DPPA 2,2-diphenylpropionic acid
  • 3DPPA 3,3-diphenylpropionic acid
  • 2,2-diphenylpropionic acid (2DPPA) or 3,3-diphenylpropionic acid (3DPPA) more preferably at least 3DPPA.
  • step (i) a carbonyl compound of general formula (I) is provided.
  • a carbonyl compound of general formula (I) is provided.
  • the residue R 1 is selected from the group consisting of branched or unbranched C2-C20-alkyl, branched or unbranched C2-C20-alkenyl, branched or unbranched C2-C20-alkinyl, branched or unbranched C1-C5-alkyl-O-C1-C5-alkyl, branched or unbranched C1-C10-alkoxy, C4-C20-cycloalkyl, C5-C20-cycloalkenyl, C5-C20-cycloalkinyl, C5-C20-aryl, C6-C20-alkylaryl, C6-C20-arylalkyl, C2-C20-heteroalkyl, C3-C20-cyclic heteroalkyl, C4-C20-heteroaryl, C5-C20-alkylheteroaryl and C5-C20-heteroarylalkyl, wherein the hetero atom
  • the residue R 1 is selected from the group consisting of branched or unbranched C2-C20-alkyl, branched or unbranched C2-C20-alkenyl, branched or unbranched C2-C20-alkinyl, branched or unbranched C1-C5-alkyl-O-C1-C5-alkyl, branched or unbranched C1-C10-alkoxy, C4-C20-cycloalkyl, C5-C20-cycloalkenyl, C5-C20-cycloalkinyl, C5-C20-aryl, C6-C20-alkylaryl, C6-C20-arylalkyl, C2-C20-heteroalkyl, C3-C20-cyclic heteroalkyl, wherein the hetero atom(s) is/are selected from oxygen, sulfur and nitrogen wherein in case of more than one aliphatic or aromatic ring system the ring systems are con
  • the residue R 1 is selected from the group consisting of branched or unbranched C2-C20-alkyl, branched or unbranched C2-C20-alkenyl, branched or unbranched C2-C20-alkinyl, C4-C20-cycloalkyl, C5-C20-cycloalkenyl, C5-C20-cycloalkinyl and C5-C20-aryl, wherein in case of more than one aliphatic or aromatic ring system the ring systems are condensed or separate, wherein each residue R 1 has at least one substituent R 1a selected from the group consisting of hydrogen atom, halogen atom, C1-C3-alkyl and C1-C3-alkoxy.
  • the residue R 1 is selected from the group consisting of branched or unbranched C2-C10-alkyl, C5-C10-cycloalkyl and C5-C20-aryl, wherein in case of more than one aliphatic or aromatic ring system the ring systems are condensed or separate, wherein each residue R 1 has at least one substituent R 1a selected from the group consisting of hydrogen atom, halogen atom, C1-C3-alkyl and C1-C3-alkoxy; R 1 being preferably selected from the group consisting of methyl, iso-propyl, cyclohexyl and phenyl, wherein phenyl has at least one substituent R 1a selected from the group consisting of hydrogen atom, halogen atom, preferably fluoro or chloro, and methoxy, preferably meta- or para-methoxy.
  • R 1 being branched or unbranched C2-C20-alkyl or branched or unbranched C2-C20-alkenyl
  • a preferred embodiment relates to R 1 being branched or unbranched C4-C20-alkyl or branched or unbranched C4-C20-alkenyl.
  • the residue R 2 is selected from the group consisting of hydrogen atom, branched or unbranched C1-C5-alkyl, wherein each residue R 2 has at least one substituent R 2a selected from the group consisting of hydrogen atom, halogen atom, C1-C3-alkyl and C1-C3-alkoxy.
  • the residue R 2 is selected from the group of branched or unbranched C1-C3-alkyl, R 2 being preferably methyl.
  • the carbonyl compound provided according to (i) is reacted in the presence of a transaminase.
  • the transaminase according to (ii) is selected from the group of transaminases, preferably from the group of amine transaminases, more preferably selected from the group consisting of amine transaminase from Aspergillus fumigates (AfATA) according to SEQ ID NO 1, amine transaminase from Gibberella zeae (GzATA) according to SEQ ID NO 2, amine transaminase from Neosartorya fischeri (NfATA) according to SEQ ID NO 3, amine transaminase from Aspergillus oryzae (AoATA) according to SEQ ID NO 4, amine transaminase from Aspergillus terreus (AtATA) according to SEQ ID NO 5, amine transaminase from Mycobacterium vanbaalenii (MvATA
  • Table 1 shows an overview of the preferred amine transaminases: Table 1: Amine transaminases of SEQ ID Nos 1 to 7 Name Abbreviation SEQ ID NO. NCBI* accession no. GenBank* accession no. amine transaminase from Aspergillus fumigates AfATA 1 XP_748821.1 amine transaminase from GzATA 2 XP_011317603.1 Gibberella zeae amine transaminase from Neosartorya fischeri NfATA 3 XP_001261640.1 amine transaminase from Aspergillus oryzae AoATA 4 BAE56564.1 amine transaminase from Aspergillus terreus AtATA 5 XP_001209325.1 amine transaminase from Mycobacterium vanbaalenii MvATA 6 WP_083062280.1 amine transaminase from Silicibacter pomeroyi SpATA 7 WP_
  • the expression "having the same function as the amine transaminase of any one of SEQ ID NO. 1 to 7” means that the homologue enzyme catalyzes an amine transaminase reaction at least with an effectivity of 90% as the transaminase of any one of SEQ ID NO. 1 to 7.
  • the homologue enzyme has a sequence identity of at least 75%, preferably at least 85%, more preferably at least 90%, more preferably at least 95%, more preferably at least 98%, more preferably at least 99%, with any one of SEQ ID NO 1 to 7 and has the same function as the amine transaminase of SEQ ID NO 1 to 7.
  • the carbonyl compound provided according to (i) is reacted in the presence of a transaminase with ii-a) at least one primary amine; and ii-b) at least one carboxylic acid.
  • the at least one primary amine according to (ii-a) is used in non-protonated form or in protonated form with a suitable counter anion.
  • the at least one primary amine according to (ii-a) is selected from the group of mono- and diamines having one to 10 carbon atoms, preferably from the group consisting of 1,5-diamino-pentane (cadaverine), alanine, 2-aminobutane (sec-butylamine) and 2-amino-propane, and is preferably 2-amino-propane (isopropylamine) in its non-protonated or protonated form, wherein the protonated form is present in combination with a suitable anion.
  • cadaverine 1,5-diamino-pentane
  • 2-aminobutane sec-butylamine
  • 2-amino-propane 2-amino-propane
  • the at least one primary amine according to (ii-a) and the at least one carboxylic acid according to (ii-b) are used as one or more salt(s) comprising the protonated form of the at least one primary amine and the deprotonated form of the at least one carboxylic acid, preferably as one salt comprising the protonated form of the at least one primary amine and the deprotonated form of the at least one carboxylic acid, more preferably as isopropyl ammonium 3,3-diphenylpropionate.
  • an amino salt compound is obtained.
  • the amino salt compound obtained according to (ii) comprises a cation of general formula (II) wherein R 1 and R 2 are as defined for general formula (I), and an anion based on the at least one carboxylic acid, which is an anion of general formula (IIIa) wherein n is zero or 1; the residues R 3 and R 4 are both phenyl or together form a phenyl ring, wherein each phenyl ring has at least one further substituent selected from the group consisting of hydrogen atom, halogen atom, preferably chloro, and nitro group and the residue R 5 is hydrogen atom or methyl or absent if R 3 and R 4 together form a phenyl ring; wherein the anion of general formula (IIIa) is preferably selected from the group consisting of diphenylacetate, 2,2-diphenylpropionate, 3,3-diphenylpropioniate, 3,4-dichloro-benzoate, 3,4
  • the reaction according to (ii) is carried out in an aqueous solution, which preferably comprises at least 80 weight-%, more preferably at least 90 weight-%, more preferably at least 95 weight-%, more preferably at least 98 weight-%, water, based on the overall weight of the aqueous solution.
  • the aqueous solution preferably comprises a buffer, preferably selected from the group of tris(hydroxymethyl)aminomethane buffer (TRIS buffer), 3-(N-morpholino)propanesulfonic acid buffer (MOPS buffer), N,N-Bis(2-hydroxyethyl)-2-aminoethanesulfonic acid buffer (BES buffer), N-(tris(hydroxymethyl)methyl)-glycine buffer (Tricine buffer), Carbonate buffer, N-cyclohexyl-2-aminoethanesulfonic acid buffer (CHES buffer), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid buffer (HEPES buffer) and phosphate buffer, more preferably at least a phosphate buffer.
  • TAS buffer tris(hydroxymethyl)aminomethane buffer
  • MOPS buffer 3-(N-morpholino)propanesulfonic acid buffer
  • BES buffer N,N-Bis(2-hydroxyethyl)-2-aminoethan
  • the reaction according to (ii) is carried out at a pH value in the range of 6.0 to 9.5, more preferably in the range of from 6.5 to 9.0, more preferably in the range of from 7.0 to 8.0.
  • the reaction according to (ii) is carried out for a time period of at least one hour, more preferably for a time in the range of from 1 to 1,000 hours, more preferably in the range of from 5 to 500 hours, more preferably in the range of from 10 to 200 hours.
  • the reaction according to (ii) is carried out at a temperature in the range of from 10 to 50 °C, more preferably in the range of from 15 to 45 °C, more preferably in the range of from 20 to 40 °C, more preferably in the range of from 25 to 35 °C.
  • the concentration of the at least one carboxylic acid is kept in the range of from 0.001 to 50 mM, preferably in the range of from 1 to 45 mM, more preferably in the range of from 5 to 40 mM during step (ii).
  • the at least one carboxylic acid being a carboxylic acid according to general formula (III), wherein the residues R 3 and R 4 are each a phenyl ring, preferably a carboxylic acid selected from the group consisting of diphenylacetic acid (DPAA), 2,2-diphenylpropionic acid (2DPPA) and 3,3-diphenylpropionic acid (3DPPA), the concentration of the at least one carboxylic acid is ⁇ 50 mM during step (ii).
  • DPAA diphenylacetic acid
  • 2DPPA 2,2-diphenylpropionic acid
  • 3DPPA 3,3-diphenylpropionic acid
  • the method comprises reaction steps (i) and (ii). According to a preferred embodiment, the method further comprises:
  • separating the crystallized amine salt compound according to (iii) from the mixture is done by sedimentation, centrifugation or filtration, preferably by filtration.
  • the method further comprises, preferably in addition to (i), (ii) and (iii):
  • the water immiscible organic solvent according to (vi) has preferably a Kow value of at least 0.5, more preferably of at least 0.6, more preferably of at least 0.7, more preferably of at least 0.8.
  • the water immiscible organic solvent according to (vi) is selected from the group of ethers, more preferably from the group of aliphatic ethers, more preferably MTBE (methyl-tert-butylether).
  • the method further comprises, preferably in addition to (i), (ii), (iii), (iv), (v) and (vi):
  • the method further comprises, preferably in addition to (i), (ii), (iii), (iv), (v) and (vi):
  • the method further comprises, preferably in addition to (i), (ii), (iii), (iv), (v), (vi) and (vii-a) or in addition to (i), (ii), (iii), (iv), (v), (vi) and (vii-b):
  • the present invention relates to an amino salt compound comprising
  • the anion of general formula (IIIa) of the amine salt compound is selected from the group consisting of diphenylacetate, 2,2-diphenylpropionate, 3,3-diphenylpropionate, 3,4-dichloro-benzoate, 3,4-dinitro-benzoate and 4-chloro-3-nitro-benzoate, more preferably from the group consisting of diphenylacetate, 2,2-diphenylpropionate, 3,3-diphenylpropionate, more preferably 2,2-diphenylpropionate or 3,3-diphenylpropionate, more preferably 3,3-diphenylpropionate.
  • the residue R 1 is selected from the group consisting of branched or unbranched C2-C20-alkyl, branched or unbranched C2-C20-alkenyl, branched or unbranched C2-C20-alkinyl, branched or unbranched C1-C5-alkyl-O-C1-C5-alkyl, branched or unbranched C1-C10-alkoxy, C4-C20-cycloalkyl, C5-C20-cycloalkenyl, C5-C20-cycloalkinyl, C5-C20-aryl, C6-C20-alkylaryl, C6-C20-arylalkyl, C2-C20-heteroalkyl, C3-C20-cyclic heteroalkyl, C4-C20-heteroaryl, C5-C20-alkylheteroaryl and C5-C20-heteroarylalkyl, wherein the hetero atom
  • the residue R 1 is selected from the group consisting of branched or unbranched C2-C20-alkyl, branched or unbranched C2-C20-alkenyl, branched or unbranched C2-C20-alkinyl, branched or unbranched C1-C5-alkyl-O-C1-C5-alkyl, branched or unbranched C1-C10-alkoxy, C4-C20-cycloalkyl, C5-C20-cycloalkenyl, C5-C20-cycloalkinyl, C5-C20-aryl, C6-C20-alkylaryl, C6-C20-arylalkyl, C2-C20-heteroalkyl, C3-C20-cyclic heteroalkyl, wherein in case of more than one aliphatic or aromatic ring system the ring systems are condensed or separate, wherein each residue R 1 has at least one substituoride, branched or unbranched C2-
  • the residue R 1 is selected from the group consisting of branched or unbranched C2-C20-alkyl, branched or unbranched C2-C20-alkenyl, branched or unbranched C2-C20-alkinyl, C4-C20-cycloalkyl, C5-C20-cycloalkenyl, C5-C20-cycloalkinyl, C5-C20-aryl, wherein in case of more than one aliphatic or aromatic ring system the ring systems are condensed or separate, wherein each residue R 1 has at least one substituent R 1a selected from the group consisting of hydrogen atom, halogen atom, C1-C3-alkyl and C1-C3-alkoxy.
  • the residue R 1 is selected from the group consisting of branched or unbranched C2-C10-alkyl, C5-C10-cycloalkyl and C5-C20-aryl, wherein in case of more than one aliphatic or aromatic ring system the ring systems are condensed or separate, wherein each residue R 1 has at least one substituent R 1a selected from the group consisting of hydrogen atom, halogen atom, C1-C3-alkyl and C1-C3-alkoxy; R 1 being preferably selected from the group consisting of methyl, iso-propyl, cyclohexyl and phenyl, wherein phenyl has at least one substituent selected from the group consisting of hydrogen atom, halogen atom, preferably fluoro or chloro, and methoxy, preferably meta- or para-methoxy.
  • R 1 being branched or unbranched C2-C20-alkyl or branched or unbranched C2-C20-alkenyl
  • a preferred embodiment relates to R 1 being branched or unbranched C4-C20-alkyl or branched or unbranched C4-C20-alkenyl.
  • the residue R 2 is selected from the group consisting of hydrogen atom, branched or unbranched C1-C5-alkyl, wherein each residue R 2 has at least one substituent R 2a selected from the group consisting of hydrogen atom, halogen atom, C1-C3-alkyl and C1-C3-alkoxy.
  • the residue R 2 is selected from the group consisting of branched or unbranched C1-C3-alkyl, R 2 being preferably methyl.
  • the combination of R 1 , R 1a being a perfluorinated alkyl is excluded.
  • R 2 and R 2a i.e. the combination of R 2 and R 2a being a perfluorinated alkyl is excluded as well.
  • neither the combination of R 1 and R 1a nor the combination of R 2 and R 2a is a perfluorinated alkyl.
  • R 1 and R 2 together form a C3-C10-cycloalkyl or a C3-C10-cycloalkenyl
  • the number of C atoms indicated includes the carbonyl C atom situated between R 1 and R 2 .
  • the present invention in another aspect relates to a composition
  • a composition comprising
  • the at least one carboxylic acid according to (b) is a carboxylic acid selected from the group consisting of diphenylacetic acid (DPAA), 2,2-diphenylpropionic acid (2DPPA), 3,3-diphenylpropionic acid (3DPPA), 3,4-dichloro-benzoic acid (34CA), 3,4-dinitro-benzoic acid (34NA) and 4-chloro-3-nitro-benzoic acid (43CNA), more preferably form the group consisting of diphenylacetic acid (DPAA), 2,2-diphenylpropionic acid (2DPPA) and 3,3-diphenylpropionic acid (3DPPA), more preferably 2,2-diphenylpropionic acid (2DPPA) or 3,3-diphenylpropionic acid (3DPPA), more preferably at least 3DPPA.
  • DPAA diphenylacetic acid
  • 2DPPA 2,2-diphenylpropionic acid
  • 3DPPA 3,3-diphenylpropionic acid
  • 3DPPA 3,
  • the residue R 1 of the amine according to (a) is selected from the group consisting of branched or unbranched C2-C20-alkyl, branched or unbranched C2-C20-alkenyl, branched or unbranched C2-C20-alkinyl, branched or unbranched C1-C5-alkyl-O-C1-C5-alkyl, branched or unbranched C1-C10-alkoxy, C4-C20-cycloalkyl, C5-C20-cycloalkenyl, C5-C20-cycloalkinyl, C5-C20-aryl, C6-C20-alkylaryl, C6-C20-arylalkyl, C2-C20-heteroalkyl, C3-C20-cyclic heteroalkyl, C4-C20-heteroaryl, C5-C20-alkylheteroaryl and C5-C20
  • the residue R 1 of the amine according to (a) is selected from the group consisting of branched or unbranched C2-C20-alkyl, branched or unbranched C2-C20-alkenyl, branched or unbranched C2-C20-alkinyl, branched or unbranched C1-C5-alkyl-O-C1-C5-alkyl, branched or unbranched C1-C10-alkoxy, C4-C20-cycloalkyl, C5-C20-cycloalkenyl, C5-C20-cycloalkinyl, C5-C20-aryl, C6-C20-alkylaryl, C6-C20-arylalkyl, C2-C20-heteroalkyl, C3-C20-cyclic heteroalkyl, wherein in case of more than one aliphatic or aromatic ring system the ring systems are condensed or separate, wherein each residue R 1 has at least one substituoride,
  • the residue R 1 of the amine according to (a) is selected from the group consisting of branched or unbranched C2-C20-alkyl, branched or unbranched C2-C20-alkenyl, branched or unbranched C2-C20-alkinyl, C4-C20-cycloalkyl, C5-C20-cycloalkenyl, and C5-C20-cycloalkinyl, C5-C20-aryl, wherein in case of more than one aliphatic or aromatic ring system the ring systems are condensed or separate, wherein each residue R 1 has at least one substituent R 1a selected from the group consisting of hydrogen atom, halogen atom, C1-C3-alkyl and C1-C3-alkoxy.
  • the residue R 1 of the amine according to (a) is selected from the group consisting of branched or unbranched C2-C10-alkyl, C5-C10-cycloalkyl and C5-C20-aryl, wherein in case of more than one aliphatic or aromatic ring system the ring systems are condensed or separate, wherein each residue R 1 has at least one substituent R 1a selected from the group consisting of hydrogen atom, halogen atom, C1-C3-alkyl and C1-C3-alkoxy; R 1 being preferably selected from the group consisting of methyl, iso-propyl, cyclohexyl and phenyl, wherein phenyl has at least one substituent selected from the group consisting of hydrogen atom, halogen atom, preferably fluoro or chloro, and methoxy, preferably meta- or para-methoxy.
  • R 1 being branched or unbranched C2-C20-alkyl or branched or unbranched C2-C20-alkenyl
  • a preferred embodiment relates to R1 being branched or unbranched C4-C20-alkyl or branched or unbranched C4-C20-alkenyl.
  • the residue R 2 of the amine according to (a) is selected from the group consisting of hydrogen atom, branched or unbranched C1-C5-alkyl, wherein each residue R 2 has at least one substituent R 2a selected from the group consisting of hydrogen atom, halogen atom, C1-C3-alkyl and C1-C3-alkoxy.
  • the residue R 2 of the amine according to (a) is selected from the group consisting of branched or unbranched C1-C3-alkyl, R 2 being preferably methyl.
  • the combination of R 1 , R 1a being a perfluorinated alkyl is excluded.
  • R 2 and R 2a i.e. the combination of R 2 and R 2a being a perfluorinated alkyl is excluded as well.
  • neither the combination of R 1 and R 1a nor the combination of R 2 and R 2a is a perfluorinated alkyl.
  • the number of C atoms indicated includes the carbonyl C atom situated between R 1 and R 2 .
  • the composition comprises the amine according to (a) in an amount in the range of from 90 to 99.9 weight-%, preferably in an amount in the range of from 95 to 99.9 weight-%, more preferably in an amount in the range of from 98 to 99.9 weight-%.
  • the composition comprises the at least one carboxylic acid according to (b) (protonated form or carboxylate with a suitable counter ion) in an amount of at least 0,003 weight-%, preferably in an amount in the range of from 0.003 to 5 weight-%, more preferably in an amount in the range of from 0.003 to 3 weight-%.
  • the straight-forward screening approach especially resulted in the identification of two benzylbenzene-based acids and three benzoic acid derivatives that matched the required criteria: DPAA, 3DPPA, 34CA, 34NA and 43CNA.
  • 2DPPA was identified as suitable carboxylic acid.
  • the isopropylamine salt of 4-chloro-3-nitrobenzoic acid (43CNA) 5 shows a very high solubility of 993 mmol ⁇ l -1
  • the 1-phenylethylamine salt of 43CNA 6a is considerable less soluble with 22 mmol ⁇ 1 -1 .
  • the solubility of amine salts can be further reduced if an over-stoichiometric amount of carboxylate is added to the mother liquor, which pushes the equilibrium from the dissociated forms (present in solution) towards its non-dissociated solid salt form.
  • product amine 2 is easily realized by dissolving product salt 6 in an aqueous solution at high pH, followed by an extraction and evaporation of the solvent, e.g. MTBE.
  • solvent e.g. MTBE.
  • the respective hydrochloride salt can be directly crystallized from the ether phase by a careful addition of concentrated HCl.
  • the spent acid 3DPPA can also be precipitated from the remaining aqueous phase by acidification with concentrated HCl, due to its low solubility at low pH.
  • ECS-ATA01 TA from Aspergillus fumigates, SEQ ID NO. 1
  • ECS-ATA02 TA from Gibberella zeae, SEQ ID NO.2
  • ECS-ATA03 TA from Neosartorya fischeri, SEQ ID NO. 3
  • ECS-ATA04 TA from Aspergillus oryzae, SEQ ID NO. 4
  • ECS-ATA05 TA from Aspergillus terreus, SEQ ID NO. 5
  • ECS-ATA07 TA from Mycobacterium vanbaalenii, SEQ IDNO. 6
  • ECS-ATA08 TA from Silicibacter pomeroyi, SEQ ID NO. 7 ).
  • HPLC Enantiomeric excess was measured with a 1100 Series HPLC by Agilent (Santa Clara, California, United States), equipped with a diode array detector, with a Chiralcel OD-H (250 mm length, 4.6 mm internal diameter, particle size: 5 ⁇ m) and a flow of 1 mL/min at 25 °C.
  • Eluent composition for the respective amine 99% n-heptane/ 1% ethanol for 2f; 98% n-heptane/ 2% ethanol for 2b, 2c and 2g; 95% n-heptane/ 5% ethanol for 2a, 2d and2e
  • Enzyme Activity Assay Enzyme activity was measured at a wave length of 245 nM with the spectrophotometer Specord 200 from Analytik Jena (Jena, Germany). Extinction coefficient of acetophenone: 11.852 (mM•cm) -1 .
  • composition of the assay 250 ⁇ L buffer solution, 250 ⁇ L 10 mM (S)-1-phenylethylamine in buffer solution, 250 ⁇ L 10 mM sodium pyruvate in buffer solution and 250 ⁇ L enzyme sample in buffer solution with 0.1 mM pyridoxal phosphate. All measurements were measured against a reference solution, whereas the enzyme solution was replaced with 200 ⁇ L buffer solution and 50 ⁇ L of 10 mM pyridoxal phosphate in buffer solution.
  • Buffer solution 50 mM phosphate buffer pH 8 with 0.25% DMSO.
  • the selected exemplary product amines were dissolved in 50 mM phosphate buffer pH 7.5 with a concentration of 100 mM each and the pH carefully adjusted back to 7.5.
  • solutions of the donor amines were prepared (200 mM for racemic 2 butylamine, racemic alanine and L-alanine and 500 mM & 2000 mM for isopropylamine and racemic 1-phenylethylamine).
  • Fig. 3 shows an exemplary result for ( S )-4-Chloro-1-phenylethylamine salts after 1 hour. The procedure was repeated for all amine solutions and the results compared against each other (table 4). The intensity of precipitation, as shown by shade code, serves as a qualitative approximation of salt solubilities and their differences.
  • Table 4 graphical presentation of the screening results exemplary donor amines exemplary product amines isopropyl-amine rac-1-phenylethylamine 2-butylamine LD-alanine L-alanine ( S )-1-PEA 2a ( S )-3Cl-1-PEA ( S )-4Cl-1-PEA ( R )-3F-1-PEA ( R )-4F-1-PEA ( S )-3MeO-1-PEA ( R )-4MeO-1-PEA 3 rac-2a 2b 2c 2d 2e 2f 2g conc.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Zoology (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Furan Compounds (AREA)
  • Pyridine Compounds (AREA)
EP17202282.4A 2017-11-17 2017-11-17 Method for preparing amines from carbonyl compounds by transaminase reaction under salt precipitation Active EP3486324B1 (en)

Priority Applications (8)

Application Number Priority Date Filing Date Title
EP17202282.4A EP3486324B1 (en) 2017-11-17 2017-11-17 Method for preparing amines from carbonyl compounds by transaminase reaction under salt precipitation
ES17202282T ES2816102T3 (es) 2017-11-17 2017-11-17 Procedimiento de preparación de aminas a partir de compuestos de carbonilo mediante reacción de transaminasa bajo precipitación de sal
DK17202282.4T DK3486324T3 (da) 2017-11-17 2017-11-17 Fremgangsmåde til fremstilling af aminer fra carbonylforbindelser ved transaminasereaktion under saltudfældning
KR1020207017054A KR102554463B1 (ko) 2017-11-17 2018-11-16 염 침전 하에 아미노전달효소 반응에 의해 카르보닐 화합물로부터 아민을 제조하는 방법
PCT/EP2018/081517 WO2019096973A1 (en) 2017-11-17 2018-11-16 Method for preparing amines from carbonyl compounds by transaminase reaction under salt precipitation
JP2020545446A JP7250807B2 (ja) 2017-11-17 2018-11-16 塩沈殿下のトランスアミナーゼ反応によりカルボニル化合物からアミンを調製する方法
US16/764,476 US12018303B2 (en) 2017-11-17 2018-11-16 Method for preparing amines from carbonyl compounds by transaminase reaction under salt precipitation
CN201880074226.2A CN111465699B (zh) 2017-11-17 2018-11-16 在盐沉淀下通过转氨酶反应由羰基化合物制备胺的方法

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
EP17202282.4A EP3486324B1 (en) 2017-11-17 2017-11-17 Method for preparing amines from carbonyl compounds by transaminase reaction under salt precipitation

Publications (2)

Publication Number Publication Date
EP3486324A1 EP3486324A1 (en) 2019-05-22
EP3486324B1 true EP3486324B1 (en) 2020-06-10

Family

ID=60654601

Family Applications (1)

Application Number Title Priority Date Filing Date
EP17202282.4A Active EP3486324B1 (en) 2017-11-17 2017-11-17 Method for preparing amines from carbonyl compounds by transaminase reaction under salt precipitation

Country Status (8)

Country Link
US (1) US12018303B2 (da)
EP (1) EP3486324B1 (da)
JP (1) JP7250807B2 (da)
KR (1) KR102554463B1 (da)
CN (1) CN111465699B (da)
DK (1) DK3486324T3 (da)
ES (1) ES2816102T3 (da)
WO (1) WO2019096973A1 (da)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3486324B1 (en) * 2017-11-17 2020-06-10 Enzymicals AG Method for preparing amines from carbonyl compounds by transaminase reaction under salt precipitation
EP4050100A4 (en) * 2019-10-25 2022-11-09 Asymchem Laboratories (Fuxin) Co., Ltd. TRANSAMINASE MUTANT AND USE THEREOF
CN111235127B (zh) * 2020-04-29 2020-08-14 凯莱英医药集团（天津）股份有限公司 转氨酶突变体及其应用
CN113881720B (zh) * 2020-12-31 2023-08-25 上海合全药物研发有限公司 一种转氨酶及使用其进行催化制备的方法
CN114277010B (zh) * 2021-12-24 2025-09-09 上海合全药物研发有限公司 转氨酶及其在制备光学纯手性胺中的应用
CN117567270B (zh) * 2023-11-20 2024-08-09 江南大学 一种醇胺-壬二酸超分子离子盐及其制备和应用

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102007042600A1 (de) 2007-09-07 2009-03-12 Evonik Degussa Gmbh Verfahren zur Herstellung von enantiomerenangereichten Aminen
SG178842A1 (en) * 2009-09-02 2012-04-27 Lonza Ag A process for the identification and preparation of a (r)-specific omega-transaminase
KR20120083908A (ko) 2009-10-13 2012-07-26 게노마티카 인코포레이티드 1,4-부탄다이올, 4-하이드록시부탄알, 4-하이드록시부티릴-ｃｏａ, 푸트레신 및 관련 화합물의 제조를 위한 미생물 및 관련 방법
ES2650245T3 (es) 2010-07-14 2018-01-17 Dpx Holdings B.V. Método enzimático para aminación (R)-selectiva
EP3486324B1 (en) * 2017-11-17 2020-06-10 Enzymicals AG Method for preparing amines from carbonyl compounds by transaminase reaction under salt precipitation

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
None *

Also Published As

Publication number Publication date
CN111465699B (zh) 2024-09-20
EP3486324A1 (en) 2019-05-22
KR20200110641A (ko) 2020-09-24
US20230167472A9 (en) 2023-06-01
DK3486324T3 (da) 2020-09-14
JP2021503306A (ja) 2021-02-12
JP7250807B2 (ja) 2023-04-03
ES2816102T3 (es) 2021-03-31
US20220162654A1 (en) 2022-05-26
WO2019096973A1 (en) 2019-05-23
US12018303B2 (en) 2024-06-25
CN111465699A (zh) 2020-07-28
KR102554463B1 (ko) 2023-07-12

Similar Documents

Publication Publication Date Title
EP3486324B1 (en) Method for preparing amines from carbonyl compounds by transaminase reaction under salt precipitation
US9255115B2 (en) Method for producing glufosinate P free acid
Mack et al. Extractive in situ product removal for the application of naturally produced L-alanine as an amine donor in enzymatic metaraminol production
Neuburger et al. Implementation and scale-up of a semi-continuous transaminase-catalyzed reactive crystallization for the preparation of (S)-(3-methoxyphenyl) ethylamine
CA2905411A1 (en) Process and intermediates for the preparation of pregabalin
RU2561602C2 (ru) Способ получения аминокислоты из 2-аминобутиролактона
US10207976B2 (en) Method for producing optically active 2-(2-fluorobiphenyl-4-yl) propanoic acid
US20160332958A1 (en) Method for preparing lacosamide
US20100256416A1 (en) Process for production of n-carbamoyl-tert-leucine
Czarnecki et al. Simple and practical direct asymmetric aldol reaction of hydroxyacetone catalyzed by 9-amino Cinchona alkaloid tartrates
JP5533654B2 (ja) ３，５−ジヒドロキシ−６−ヘプテン酸アミン塩類の製造方法
US20200377456A1 (en) Process for the preparation of glycopyrrolate tosylate
Labib et al. Metabolic and Process Engineering for Microbial Production of Protocatechuate from Xylose with Corynebacterium glutamicum
Pankaskie et al. An Improved Synthetic Route to Aminoxypropylamine (Apa) and Related Homologs
WO2007119302A1 (ja) アミノ酸リン酸類塩の製造方法
EP3246308A1 (en) Method for producing aromatic amine compound
JP3847934B2 (ja) γ−オキソ−ホモフェニルアラニン誘導体及びそれを還元してなるホモフェニルアラニン誘導体の製造方法
EP0302624A2 (en) A method for racemization of optically active serine
Chen et al. A synthesis of N 6, N 6, N 6-trimethyl-L-lysine dioxalate in gram amounts
EP3348543B1 (en) Method for preparing d-arginine
JP2011057619A (ja) 光学活性アミン化合物の製造方法、並びに、ジアステレオマー塩及びその製造方法
KR0152258B1 (ko) (-)2-치환 오르니틴의 제조방법
Labib et al. Metabolic and process engineering for microbial production of
KR20160022618A (ko) D-아르기닌의 제조 방법
WO2012033187A1 (ja) 光学活性２－アミノ酪酸の製造方法

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION HAS BEEN PUBLISHED

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

AX Request for extension of the european patent

Extension state: BA ME

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: REQUEST FOR EXAMINATION WAS MADE

17P Request for examination filed

Effective date: 20191121

GRAP Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOSNIGR1

RBV Designated contracting states (corrected)

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: GRANT OF PATENT IS INTENDED

INTG Intention to grant announced

Effective date: 20200102

GRAS Grant fee paid

Free format text: ORIGINAL CODE: EPIDOSNIGR3

GRAA (expected) grant

Free format text: ORIGINAL CODE: 0009210

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE PATENT HAS BEEN GRANTED

AK Designated contracting states

Kind code of ref document: B1

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

REG Reference to a national code

Ref country code: GB

Ref legal event code: FG4D

REG Reference to a national code

Ref country code: CH

Ref legal event code: EP

Ref country code: AT

Ref legal event code: REF

Ref document number: 1279232

Country of ref document: AT

Kind code of ref document: T

Effective date: 20200615

REG Reference to a national code

Ref country code: DE

Ref legal event code: R096

Ref document number: 602017017924

Country of ref document: DE

REG Reference to a national code

Ref country code: IE

Ref legal event code: FG4D

REG Reference to a national code

Ref country code: CH

Ref legal event code: NV

Representative=s name: SCHMAUDER AND PARTNER AG PATENT- UND MARKENANW, CH

REG Reference to a national code

Ref country code: DK

Ref legal event code: T3

Effective date: 20200909

REG Reference to a national code

Ref country code: NL

Ref legal event code: FP

REG Reference to a national code

Ref country code: LT

Ref legal event code: MG4D

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: NO

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200910

Ref country code: LT

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200610

Ref country code: FI

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200610

Ref country code: SE

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200610

Ref country code: GR

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200911

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: BG

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200910

Ref country code: RS

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200610

Ref country code: LV

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200610

Ref country code: HR

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200610

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: AL

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200610

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: SM

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200610

Ref country code: EE

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200610

Ref country code: CZ

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200610

Ref country code: RO

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200610

Ref country code: PT

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20201012

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: PL

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200610

Ref country code: SK

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200610

Ref country code: IS

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20201010

REG Reference to a national code

Ref country code: DE

Ref legal event code: R097

Ref document number: 602017017924

Country of ref document: DE

REG Reference to a national code

Ref country code: ES

Ref legal event code: FG2A

Ref document number: 2816102

Country of ref document: ES

Kind code of ref document: T3

Effective date: 20210331

PLBE No opposition filed within time limit

Free format text: ORIGINAL CODE: 0009261

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT

26N No opposition filed

Effective date: 20210311

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: SI

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200610

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: MC

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200610

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: LU

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20201117

REG Reference to a national code

Ref country code: BE

Ref legal event code: MM

Effective date: 20201130

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: IE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20201117

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: TR

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200610

Ref country code: MT

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200610

Ref country code: CY

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200610

REG Reference to a national code

Ref country code: AT

Ref legal event code: UEP

Ref document number: 1279232

Country of ref document: AT

Kind code of ref document: T

Effective date: 20200610

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: MK

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20200610

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: BE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20201130

REG Reference to a national code

Ref country code: CH

Ref legal event code: U11

Free format text: ST27 STATUS EVENT CODE: U-0-0-U10-U11 (AS PROVIDED BY THE NATIONAL OFFICE)

Effective date: 20251201

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: NL

Payment date: 20251119

Year of fee payment: 9

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: DE

Payment date: 20251118

Year of fee payment: 9

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: GB

Payment date: 20251120

Year of fee payment: 9

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: AT

Payment date: 20251117

Year of fee payment: 9

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: IT

Payment date: 20251128

Year of fee payment: 9

Ref country code: DK

Payment date: 20251119

Year of fee payment: 9

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: FR

Payment date: 20251120

Year of fee payment: 9

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: CH

Payment date: 20251201

Year of fee payment: 9

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: ES

Payment date: 20251216

Year of fee payment: 9